SVIBOR - Project code: 1-08-271

MINISTRY OF SCIENCE AND TECHNOLOGY

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Project code: 1-08-271


Detection of mutations in chromosomal DNA using virus vectors


Main researcher: NAGY, BISERKA (33076)


Assistants
Type of research: basic
Duration from: 01/01/91. to 12/31/95.
Papers on project (total): 22
Papers on project quoted in Current Contents: 4
Institution name: Prirodoslovno-matematički fakultet, Prirodoslovni odjeli, Zagreb (119)
Department/Institute: BIOLOGICAL DIVISION DEPARTMENT OF MOLECULAR BIOLOGY
Address: Rooseveltov trg 6
City: 10000 - Zagreb, Croatia
Communication
Phone: 385 (0)1 4552 646
Phone: 385 (0)1 442 804
Phone: 385 (0)1 442 604
Fax: 385 (0)1 4552 645
Summary: Radiation and chemicaly induced mutagenesis in mammalian cells has been studied for sometime, the molecular mechanisam responsible for this effect remain obscured. While repair following mutagenic treatment is thought to play an especially important role in the mutagenic process, such studies are limited. The preliminary goal of this proposal is to determine specifically the significance and the effects of DNA polymerization on mutation rate and lesion spectrum.The process is integral parts of DNA replication and is believed to be essential to maintain DNA fidelity. Studies ytilize the retroviral shuttle vector pZipGptNeo to determine the specificity of spontaneous and radiation and chemicaly induced mutations. The construction of a new retroviral shuttle vector and its introduction into repair deficient and proficient CHO cell lines will be included. Preliminary experiments suggest that gpt gene mutations of shuttle vector induced into mouse L cells are similar tu HGPRT mutations in CHO cells . A research plan has been formulated to further elucidate these special repair processes.

Keywords: Mutational specificity, plasmids, virus vectors, mutagenic agens

Research goals: Thepurpose of this project is to determine the mutational eventsin eukariotic cells. Many chemicals as well as radiation are theknown mutagen and carcinogen and exposure to this agent canoccure in the envirnoment. The determination of the specifity ofmutagenesis should provide important clues to the mechanism bywhich those agents produces mutations and cancer. Thedetermination of the actual base changes whih occur when a geneis mutated is an important step in chicidating the detailedmechanisms of mutagenesis. Several systems have been describedwhich this make possible. In the shuttle vector systems, a target gene is introduced intothe mammalian cells. Following mutagenesis in the mammalian cells environment, theshuttle vector sequences are recovered and introduced intobacteria. In the bacteria, large amounts of the mutant genesequence can be produced for DNA base sequence analysis. Clearly,shuttle vector systems have become on important tool in the studyof mutational mechanisms in higher organisms. A mouse fibroblast cell line was constracted in wich a retroviral shuttle vector, capable of replication in both mammalian cells and bacteria.Retroviral shuttle vector contains E.coli gpt gen.The cells are treated with mutagens and cell line having mutations in gpt gene will be isolated.A number of agents andtreatment conditions are known to modulate the biological effects of radiation.We will determine what conditions modulate the frequencz of gpt mutants induced in our cell lines.

COOPERATION - PROJECTS


  1. Name of project: > GENES AND ENZYMES INVOLVED IN THE INHIBITION OF RECOMBINATION
    Name of institution: INTERNATIONAL CENTRE FOR GENETIC ENGINNERING AND BIOTECHNOLOGY
    City: Trieste, Italy

  2. Name of project: > MOLECULAR ASPECTS OF RADIATION CARCINOGENESIS
    Name of institution: UNIVERSITY OF CHICAGO, DEPT. OF RADIATION AND CELLULAR ONCOLOGY
    City: Chicago, Illinois, USA

COOPERATION - INSTITUTIONS


  1. Name of institution: INTERNATIONAL SCHOOL FOR ADVANCED STUDY
    Type of institution: University/Faculty
    Type of cooperation: Systematic exchange of experts
    City: Trieste, Italy

  2. Name of institution: UNIVERSITY OF CHICAGO, DEPT. OF RADIATION AND CELLULAR ONCOLOGY
    Type of institution: University/Faculty
    Type of cooperation: Occasional exchange of experts
    City: Chicago, Illinois, USA

  3. Name of institution: CANCEER RESEARCH CENTER, LAYOLLA CANCER RESEARCH FOUNDATION,
    Type of institution: University/Faculty
    Type of cooperation: Occasional exchange of experts
    City: LA YOLLA, CALIFORNIA, USA

OTHER ACHIEVEMENTS


  1. Name: IN SITZ HZBRIDISATION OF BRAIN SECTIONS OF NEONATAL AND ADULT RATS USING P1 AND P2 GABA RECEPTOR SUBUNIT SPECIFIC OLIGONUCLEOTIDE PROBES.THE autorradiographic signals obtained with p1+p2 probe in the neonatal brains were surprisinglz strong.
    Type of achievement: Technology
    Authors: Miranda Mladinić, Enrico Cherubini,
Other information about the project.
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